Methods for enhancing the degradation or conversion of cellulosic material

Abstract

The present invention relates to methods for degrading or converting a cellulosic material and for producing a substance from a cellulosic material.

1. A method for degrading or converting a cellulosic material, comprising: treating the cellulosic material with an effective amount of one or more cellulolytic proteins in the presence of an effective amount of a polypeptide having cellulolytic enhancing activity, wherein the presence of the polypeptide having cellulolytic enhancing activity increases the degradation of cellulosic material compared to the absence of the polypeptide having cellulolytic enhancing activity and wherein the polypeptide having cellulolytic enhancing activity does not itself degrade or convert the cellulosic material in the absence of one or more cellulolytic protein and wherein the polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
 (a) a polypeptide comprising an amino acid sequence which has at least 95% sequence identity with the mature polypeptide of SEQ ID NO: 2; 
 (b) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence which has at least 95% sequence identity with the mature polypeptide coding sequence of SEQ ID NO: 1; and 
 (c) a polypeptide encoded by a polynucleotide which hybridizes under at least high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the genomic DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO: 1, or (iii) a full-length complementary strand of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C. 
 
     
     
       2. The method of  claim 1 , further comprising treating the cellulosic material with an effective amount of one or more enzymes selected from the group consisting of a hemicellulase, esterase, protease, laccase, peroxidase, or a mixture thereof. 
     
     
       3. The method of  claim 1 , further comprising recovering the degraded cellulosic material. 
     
     
       4. The method of  claim 1 , wherein the cellulolytic protein and/or the polypeptide having cellulolytic enhancing activity are in the form of a fermentation broth with or without cells. 
     
     
       5. The method of  claim 1 , wherein the polypeptide having cellulolytic enhancing activity comprises or consists of the amino acid, sequence of SEQ ID NO: 2 or the mature polypeptide thereof; or a fragment thereof having cellulolytic enhancing activity. 
     
     
       6. The method of  claim 1 , wherein the polypeptide having cellulolytic enhancing activity is encoded by the polynucleotide contained in plasmid pTr3337 which is contained in  E. coli  NRRL B-30878. 
     
     
       7. The method of  claim 1 , wherein the mature polypeptide coding sequence of SEQ ID NO: 1 is nucleotides 77 to 766 of SEQ ID NO: 1. 
     
     
       8. The method of  claims 1 , wherein the mature polypeptide of SEQ ID NO: 2 is amino acids 20 to 249 of SEQ ID NO: 2. 
     
     
       9. The method of  claim 1 , wherein the polypeptide having cellulolytic enhancing activity comprises an amino acid sequence which has at least 95% sequence identity with the mature polypeptide of SEQ ID NO: 2. 
     
     
       10. The method of  claim 9 , wherein the polypeptide having cellulolytic enhancing activity comprises an amino acid sequence which has at least 97% sequence identity with the mature polypeptide of SEQ ID NO: 2. 
     
     
       11. The method of  claim 1 , wherein the polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide comprising a nucleotide sequence which has at least 95% a sequence identity with the mature polypeptide coding sequence of SEQ ID NO: 1. 
     
     
       12. The method of  claim 11 , wherein the polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide comprising a nucleotide sequence which has at least 97% a sequence identity with the mature polypeptide coding sequence of SEQ ID NO: 1. 
     
     
       13. The method of  claim 1 , wherein the polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide which hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the genomic DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO: 1, or (iii) a full-length complementary strand of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.30% SDS, 200 μg/ml sheared and denatured salmon sperm DNA and 50% a formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.